Summary
The role of exocytosis in the cellular defence reactions of arthropods was investigated using in vitro cultures of isolated haemocytes (blood cells) from the freshwater crayfish Pacifastacus leniusculus, and the shore crab Carcinus maenas. In both species, activated lysates of those cell types that contain the prophenoloxidase activating system (granular cells of crab and crayfish and semigranular cells of crayfish) were found to induce degranulation (exocytosis) of semigranular and granular cells. A cell lysate, in which the prophenoloxidase system was kept inactive, did not have this effect. Limited degranulation of granular cells of crab was also induced by lipopolysaccharides as has earlier been shown for crayfish semigranular cells. The phagocytic capability of semigranular cells from crayfish was lost after exocytosis induced by the Ca2+ ionophore A23187, and under no conditions were the granular cells of crabs or crayfish seen to ingest bacteria in vitro. An opsonic function for the attaching proteins of a β1,3-glucan-activated haemocyte lysate was demonstrated using the phagocytic hyaline cells from crabs. Phenoloxidase appeared to lack opsonic properties.
We suggest that, in crustaceans, opsonization takes place through hierarchically stimulated exocytotic release, and biochemical activation of the prophenoloxidase activating system: first from lipopolysaccharide-sensitive cells (semigranular cells of crayfish or granular cells of crabs) and then from granular cells, triggered by the initially released and activated prophenoloxidase system. Finally, “sticky” proteins of the activated prophenoloxidase system coat the invader, rendering it susceptible to the phagocytes (hyaline cells in both crab and crayfish and, to a lesser extent, semigranular cells of crayfish). These processes would, together, constitute a cellular communication pathway not previously demonstrated for invertebrates.
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Abbreviations
- DMSO:
-
dimethyl sulfoxide
- L-DOPA:
-
L-dihydroxy-phenylalanine
- GLS:
-
granular cell lysate supernatant
- HLS:
-
haemocyte lysate supernatant
- HyLS:
-
hyaline cell lysate supernatant
- LPS:
-
lipopolysaccharide
- proPO:
-
prophenoloxidase
- SGLS:
-
semigranular cell lysate supernatant
- SITS:
-
4-acetamido-4′-isothiocyanatostilbene-2,2′-disulfonic acid disodium salt
References
Ashida M, Söderhäll K (1984) The prophenoloxidase activating system in crayfish. Comp Biochem Physiol 77B: 21–26
Ashida M, Ishizaki Y, Iwahana H (1983) Activation of pro-phenoloxidase by bacterial cell walls or by β1,3-glucans in plasma of the silkworm, Bombyx mori. Biochem Biophys Res Commun 113:562–568
Götz P, Boman HG (1985) Insect immunity. In: GA Kerkut, LI Gilbert (eds) Comprehensive insect physiology, biochemistry and pharmacology. Pergamon Press, Oxford, pp 453–485
Hergenhahn H-G, Söderhäll K (1985) α2-Macroglobulin-like activity in plasma of the crayfish Pacifastacus leniusculus. Comp Biochem Physiol 818:833–835
Johansson MW, Söderhäll K (1985) Exocytosis of the prophenoloxidase activating system from crayfish haemocytes. J Comp Physiol 156B: 175–181
Leonard C, Söderhäll K, Ratcliffe NA (1985a) Studies on prophenoloxidase and protease activity of Blaberus craniifer haemocytes. Insect Biochem 15:803–810
Leonard C, Ratcliffe NA, Rowley AF (1985b) The role of prophenoloxidase activation in non-self recognition and phagocytosis by insect blood cells. J Insect Physiol 31:789–799
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Ratcliffe NA, Gagen SJ (1977) Studies on the in vivo cellular reactions of insects: an ultrastructural analysis of nodule formaiton in Galleria mellonella. Tissue Cell 9:73–85
Ratcliffe NA, White KN, Rowley AF, Walters JB (1982) Cellular defence systems of the Arthropoda. In: N Cohen, M Sigel (eds) The Reticuloendothelial System. A Comparative Treatisc Vol 3. Plenum Press, New York, pp 167–255
Ratcliffe NA, Leonard C, Rowley AF (1984) Prophenoloxidase activation: nonself recognition and cell cooperation in insect immunity. Science 226:557–559
Salt G (1970) The cellular defence reactions of insects. Cambridge University, London New York
Smith VJ, Ratcliffe NA (1978) Host defence reactions of the shore crab Carcinus maenas (L.) in vitro. J Mar Biol Assoc UK 58:367–379
Smith VJ, Söderhäll K (1983a) β-1,3 glucan activation of crustacean hemocytes in vitro and in vivo. Biol Bull (Woods Hole) 164:299–314
Smith VJ, Söderhäll K (1983b) Induction of degranulation and lysis of haemocytes in the freshwater crayfish, Astacus astacus by components of the prophenoloxidase activating system in vitro. Cell Tiss Res 233:295–303
Smith VJ, Söderhäll K, Hamilton M (1984) β1,3-glucan induced cellular defence reactions in the shore crab Carcinus maenas. Comp Biochem Physiol 77A: 635–639
Söderhäll K (1981) Fungal cell wall β1,3-glucans induce clotting and phenoloxidase attachment to foreign surfaces of crayfish hemocyte lysate. Dev Comp Immunol 5:565–573
Söderhäll K (1982) Prophenoloxidase activating system and melanization — a recognition mechanism of arthropods? A review. Dev Comp Immunol 6:601–611
Söderhäll K, Ajaxon R (1982) Effect of quinones and melanin on mycelial growth of Aphanomyces spp and extracellular protease of Aphanomvces astaci, a parasite on crayfish. J Invert Pathol 39:105–109
Söderhäll K, Häll L (1984) Lipopolysaccharide-induced activation of prophenoloxidase activating system in crayfish haemocyte lysate. Biochim Biophys Acta 797:99–104
Söderhäll K, Smith VJ (1983) Separation of the haemocyte populations of Carcinus maenas and other marine decapods, and prophenoloxidase distribution. Dev Comp Immunol 7:229–239
Söderhäll K, Smith VJ (1984) The prophenoloxidase activating system — a complement like pathway in arthropods? In: Roberts DW, Aist JR (eds) Infection processes of fungi. The Rockefeller Foundation, New York, pp 160–167
Söderhäll K, Smith VJ (1986) The prophenoloxidase activating system: the biochemistry of its activation and role in arthropod cellular immunity with special reference to crustaceans. In: Bréhélin M (ed) Immunity in invertebrates, Springer, Berlin Heidelberg New York Tokyo pp 208–223
Söderhäll K, Unestam T (1979) Activation of crayfish serum prophenol-oxidase: the specificity of cell wall glucan activation and activation by purified fungal glycoproteins of crayfish phenoloxidase. Can J Microbiol 25:406–414
Söderhäll K, Vey A, Ramstedt M (1984) Hemocyte lysate enhancement of fungal spore encapsulation by crayfish hemocytes. Dev Comp Immunol 8:23–29
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Söderhäll, K., Smith, V.J. & Johansson, M.W. Exocytosis and uptake of bacteria by isolated haemocyte populations of two crustaceans: evidence for cellular co-operation in the defence reactions of arthropods. Cell Tissue Res. 245, 43–49 (1986). https://doi.org/10.1007/BF00218085
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DOI: https://doi.org/10.1007/BF00218085